摘要
目的初步探讨金雀异黄素(genistein)与抗Ⅳ型胶原抗体耦联的方法。方法采用活性酯法将抗Ⅳ型胶原抗体与金雀异黄素进行耦联;采用凝胶层析对耦联物提纯;紫外吸收光谱法和Bradford蛋白检测法检测耦联物中的金雀异黄素与抗Ⅳ型胶原抗体的克分子比;免疫组织化学法检测耦联物与人晶状体囊膜结合的特异性。结果抗Ⅳ型胶原抗体与金雀异黄素成功耦联,耦联产物中抗Ⅳ型胶原抗体与金雀异黄素克分子比为1∶439,耦联物与晶状体囊膜可特异结合。结论采用活性酯法可成功地将抗Ⅳ型胶原抗体与金雀异黄素耦联,耦联物的免疫活性良好。
Objective To generate an immunoconjugate which genistein could be coupled to anti-collagen monoclonal antibody. Methods Genistein was bound to anti-collagen monoclonal antibody by the active ester intermediate. The immunoconjugate was purified by sephadex G25 column chromatography. The molar ratio of genistein and monoclonal antibody in the conjugate was measured by Bradford protein assay and UV absorption spectra assay. The immunological activity of the antibody in conjugate was determined by immunohistochemistry method. Results Genistein could be successfully coupled with anticollagen Ⅳ monoclonal antibody. The mass concentration of genistein in immunoconjugate was 0. 075 7 mg/ml, the mass concentration of IgG was 0. 102 mg/ml. The molar radio of genistein to monoclonal antibody in the conjugate was 439：1 ,showing that each monoclonal antibody could carry 439 drug moleculars in the conjugate. The double curve of immunoconjugate by sephadex G25 was very similar,which suggested the effect of conjoining was better. The immunoconjugate could specially bind to lens capsule. Conclusion Genistein could be coupled to anti-collagen monoclonal antibody by the active ester intermediate.
出处
《眼科研究》
CSCD
北大核心
2005年第5期453-455,共3页
Chinese Ophthalmic Research
基金
国家教育部归国留学人员基金资助(2002247)
